Protein Disulfide Isomerase Is Cleaved by Caspase-3 and -7 during Apoptosis

  • Na, Kyung Sook (Translational Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Park, Byoung Chul (Translational Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Jang, Mi (Translational Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Cho, Sayeon (Department of Pharmacy, Chung-Ang University) ;
  • Lee, Do Hee (Translational Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kang, Sunghyun (Translational Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Lee, Chong-Kil (Department of Pharmacy, Chungbuk National University) ;
  • Bae, Kwang-Hee (Translational Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Park, Sung Goo (Translational Research Center, Korea Research Institute of Bioscience and Biotechnology)
  • Received : 2007.04.12
  • Accepted : 2007.05.23
  • Published : 2007.10.31


Apoptotic signals are typically accompanied by activation of aspartate-specific cysteine proteases called caspases, and caspase-3 and -7 play crucial roles in the execution of apoptosis. Previously, using the proteomic approach, protein disulfide isomerase (PDI) was found to be a candidate substrate of caspase-7. This abundant 55 kDa protein introduces disulfide bonds into proteins (via its oxidase activity) and catalyzes the rearrangement of incorrect disulfide bonds (via its isomerase activity). PDI is abundant in the ER but is also found in non-ER locations. In this study we demonstrated that PDI is cleaved by caspase-3 and -7 in vitro. In addition, in vivo experiment showed that it is cleaved during etoposide-induced apoptosis in HL-60 cells. Subcellular fractionation showed that PDI was also present in the cytosol. Furthermore, only cytosolic PDI was clearly digested by caspase-3 and -7. It was also confirmed by confocal image analysis that PDI and caspase-7 partially co-localize in both resting and apoptotic MCF-7 cells. Overexpression of cytosolic PDI (ER retention sequence deleted) inhibited cell death after an apoptotic stimulus. These data indicate that cytosolic PDI is a substrate of caspase-3 and -7, and that it has an anti-apoptotic action.


Supported by : Korea Science & Engineering Foundation, Ministry of Health & Welfare


  1. Higuchi, T., Watanabe, Y., and Waga, I. (2004) Protein disulfide isomerase suppresses the transcriptional activity of NF-${\kappa}B$. Biochem. Biophys. Res. Commun. 318, 46−52
  2. Kang, S., Kim, E. Y., Bahn, Y. J., Chung, J. W., Lee, D. H., et al. (2007) A proteomic analysis of the effect of MAPK pathway activation on L-glutamate-induced neuronal cell death. Cell. Mol. Biol. Lett. 12, 139−147
  3. Lee, M. S. and Kim, Y. J. (2007) Pattern-recognition receptor signaling initiated from extracellular, membrane, and cytoplasmic space. Mol. Cells 23, 1−10
  4. Nicholson, D. W. (1999) Caspase structure, proteolytic substrates, and function during apoptotic cell death. Cell Death Differ. 6, 1028−1042
  5. Park, B., Lee, S., Kim, E., Cho, K., Riddell, S. R., et al. (2006) Redox regulation facilitates optimal peptide selection by MHC class I during antigen processing. Cell 127, 369−382
  6. Reddy, R. K., Mao, C., Baumeister, P., Austin, R. C., Kaufman, R. J., et al. (2003) Endoplasmic reticulum chaperone protein GRP78 protects cells from apoptosis induced by topoisomerase inhibitors: role of ATP binding site in suppression of caspase-7 activation. J. Biol. Chem. 278, 20915−20924
  7. Timmer, J. C. and Salvesen, G. S. (2007) Caspase substrates. Cell Death Differ. 14, 66−72
  8. Wilkinson, B. and Gilbert, H. F. (2004) Protein disulfide isomerase. Biochim. Biophys. Acta 1699, 35−44
  9. Yu, S. W., Wang, H., Poitras, M. F., Coombs, C., Bowers, W. J., et al. (2002) Mediation of poly(ADP-ribose) polymerase-1- dependent cell death by apoptosis-inducing factor. Science 297, 259−263
  10. Zhivotovsky, B., Samali, A., Gahm, A., and Orrenius, S. (1999) Caspases: their intracellular localization and translocation during apoptosis. Cell Death Differ. 6, 644−651
  11. Turano, C., Coppari, S., Altieri, F., and Ferraro, A. (2002) Proteins of the PDI family: unpredicted non-ER locations and functions. J. Cell Physiol. 193,154−163
  12. Fuentes-Prior, P. and Salvesen, G. S. (2004) The protein structures that shape caspase activity, specificity, activation and inhibition. Biochem. J. 384, 201−232
  13. Lee, A. Y., Park, B. C., Jang, M., Cho, S., Lee, D. H., et al. (2004) Identification of caspase-3 degradome by two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionizationtime of flight analysis. Proteomics 4, 3429−3436
  14. Dahmer, M. K. (2005) Caspases-2, -3, and -7 are involved in thapsigargin-induced apoptosis of SH-SY5Y neuroblastoma cells. J. Neurosci. Res. 80, 576−583
  15. Kulp, M. S., Frickel, E. M., Ellgaard, L., and Weissman, J. S. (2006) Domain architecture of protein-disulfide isomerase facilitates its dual role as an oxidase and an isomerase in Ero1p-mediated disulfide formation. J. Biol. Chem. 281, 876− 884
  16. Kumar, S. (2007) Caspase function in programmed cell death. Cell Death Differ. 14, 32−43
  17. Stennicke, H. R. and Salvesen, G. S. (2000) Caspases - controlling intracellular signals by protease zymogen activation. Biochim. Biophys. Acta 1477, 299−306
  18. Jessop, C. E., Chakravarthi, S., Watkins, R. H., and Bulleid, N. J. (2004) Oxidative protein folding in the mammalian endoplasmic reticulum. Biochem. Soc. Trans. 32, 655−658
  19. Jordan, P. A. and Gibbins, J. M. (2006) Extracellular disulfide exchange and the regulation of cellular function. Antioxi. Redox Signal 8, 312−324
  20. Tian, R., Li, S. J., Wang, D. L., Zhao, Z., Liu, Y., et al. (2004) The acidic C-terminal domain stabilizes the chaperone function of protein disulfide isomerase. J. Biol. Chem. 279, 48830− 48835
  21. Tanaka, S., Uehara, T., and Nomura, Y. (2000) Up-regulation of protein disulfide isomerase in response to hypoxia/brain ischemia and its protective effect against apoptotic cell death. J. Biol. Chem. 275, 10388−10393
  22. Sun, F. C., Wei, S., Li, C. W., Chang, Y. S., Chao, C. C., et al. (2006) Localization of GRP78 to mitochondria under the unfolded protein response. Biochem. J. 396, 31−39
  23. Coppari, S., Altieri, F., Ferraro, A., Chichiarelli, S., Eufemi, M., et al. (2002) Nuclear localization and DNA interaction of protein disulfide isomerase ERp57 in mammalian cells. J. Cell. Biochem. 85, 325−333
  24. Jang, M., Park, B. C., Lee, A. Y., Na, K. S., Kang, S., et al. (2007) Caspase-7 dependent cleavage of proteasome subunits during apoptosis. Biochem. Biophys. Res. Commun. Doi: 10.1016/:bbrc. 2007.08.183
  25. Takizawa, T., Tatematsu, C., Watanabe, K., Kato, K., and Nakanishi, Y. (2004) Cleavage of calnexin caused by apoptotic stimuli: implication for the regulation of apoptosis. J. Biochem. 136, 399−405
  26. Goplen, D., Wang, J., Enger, P., Tysnes, B. B., Terzis, A. J. A., et al. (2006) Protein disulfide isomerase expression is related to the invasive properties of malignant glioma. Cancer Res. 66, 9895−9902